ANTAGONISTS OF Bcl-2 AND USES THEREOF IN INDUCTION OF APOPTOSIS

Abstract

The present invention provides an antagonist of a Bcl-2 pro survival protein containing a BH3-like domain. The antagonist of the invention comprises ARTS and any fragment or peptide that comprises a BH3-like domain. The invention further provides compositions, combined compositions and kits as well as methods for treating Bcl-2 over-expressing disorders.

1 . An antagonist of a B-cell lymphoma 2 (Bcl-2) prosurvival protein comprising Apoptosis Related Protein in the TGF-beta Signaling Pathway (ARTS) and any fragment, peptide, analogues and derivatives thereof, wherein said fragment or peptide of ARTS comprises a Bcl-2 homology domain 3 (BH3)-like domain. 
     
     
         2 . The antagonist according to  claim 1 , wherein said fragment or peptide comprises the amino acid sequence of any one of residues 1-128, 1-148, 106-148, 106-128, 106-133, 112-148, 112-128, 112-133, 106-140 and 112-126 of ARTS and wherein said ARTS fragment or peptide comprises an amino acid sequence as denoted by any one of SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 35 and SEQ ID NO: 36, respectively. 
     
     
         3 . (canceled) 
     
     
         4 . The antagonist according to  claim 1 , wherein said Bcl-2 prosurvival protein is at least one of Bcl-2, Bcl-xL, Mcl-1, Bcl-w, A1/Bfl-1 and Bcl-B/Bcl2L10. 
     
     
         5 . The antagonist according to  claim 1 , wherein at least one of:
 (a) said antagonist mediates ubiquitin proteasome system (UPS) degradation of said Bcl-2 prosurvival protein, thereby enhancing or inducing apoptosis and   (b) said antagonist interacts with at least one pro-apoptotic protein member of the Bcl-2 family, thereby enhancing apoptosis.   
     
     
         6 . (canceled) 
     
     
         7 . A composition comprising an effective amount of at least one antagonist of a Bcl-2 prosurvival protein according to  claim 1 , said antagonist comprises ARTS or any fragment, peptide, analogues and derivatives thereof, wherein said fragment or peptide of ARTS comprises a BH3-like domain. 
     
     
         8 - 12 . (canceled) 
     
     
         13 . The composition according to  claim 7 , wherein said composition is a pharmaceutical composition for treating, inhibiting, preventing, ameliorating or delaying the onset of a Bcl-2 over-expressing pathological disorder, said composition optionally further comprises at least one pharmaceutically acceptable carrier, diluent or excipient. 
     
     
         14 . (canceled) 
     
     
         15 . The pharmaceutical composition according to  claim 7 , wherein said composition is adapted for use before, simultaneously with, after or any combination thereof any one of at least one BH3 mimetics compound, at least one pro-apoptotic protein member of the Bcl-2 family or any combinations thereof. 
     
     
         16 . The pharmaceutical composition according to  claim 7 , wherein said composition further comprises any one of at least one BH3 mimetics compound at least one pro-apoptotic protein member of the Bcl-2 family or any combinations thereof, wherein said BH3 mimetics compound/antagonist is at least one of 4-[4-[[2-(4-Chlorophenyl)-5,5-dimethyl-1-cyclohexen-1-yl]methyl]-1-piperazinyl]-N-[[4-[[(1R)-3-(4-morpholinyl)-1-[(phenylthio)methyl]propyl]amino]-3-[(trifluoromethyl)sulfonyl]phenyl]sulfonyl]benzamide (ABT-263), (R)-4-(4-((4′-chloro-1,1′-biphenyl]-2-yl)methyl)piperazin-1-yl)-N-((4-((4-(dimethylamino)-1-(phenylthio)butan-2-yl)amino)-3-nitrophenyl)sulfonyl)benzamide (ABT-737), 1,1′,6,6′,7,7′-hexahydroxy-5,5′-diisopropyl-3,3′-dimethyl-[2,2′-binaphthalene]-8,8′-dicarbaldehyde (AT-101), (Z)-2-(2-((3,5-dimethyl-1H-pyrrol-2-yl)methylene)-3-methoxy-2H-pyrrol-5-yl)-1H-indole methanesulfonate (GX15-070), or DNA, d(P-thio)(T-C-T-C-C-C-A-G-C-G-T-G-C-G-C-C-A-T) (oblimersen sodium) and wherein said pro-apoptotic protein member of the Bcl-2 family is any one of Bax, Bak, Bnip3, Nix/Bnip3L, Bid, Noxa, Puma and Bad. 
     
     
         17 . The pharmaceutical composition according to  claim 15 , wherein said BH3 mimetics compound/antagonist is at least one of 4-[4-[[2-(4-Chlorophenyl)-5,5-dimethyl-1-cyclohexen-1-yl]methyl]-1-piperazinyl]-N-[[4-[[(1R)-3-(4-morpholinyl)-1-[(phenylthio)methyl]propyl]amino]-3-[(trifluoromethyl)sulfonyl]phenyl]sulfonyl]benzamide (ABT-263), (R)-4-(4-((4′-chloro-[1,1′-biphenyl]-2-yl)methyl)piperazin-1-yl)-N-((4-((4-(dimethylamino)-1-(phenylthio)butan-2-yl)amino)-3-nitrophenyl)sulfonyl)benzamide (ABT-737), 1,1′,6,6′,7,7′-hexahydroxy-5,5′-diisopropyl-3,3′-dimethyl-[2,2′-binaphthalene]-8,8′-dicarbaldehyde (AT-101), (Z)-2-(2-((3,5-dimethyl-1H-pyrrol-2-yl)methylene)-3-methoxy-2H-pyrrol-5-yl)-1H-indole methanesulfonate (GX15-070), or DNA, d(P-thio)(T-C-T-C-C-C-A-G-C-G-T-G-C-G-C-C-A-T) (oblimersen sodium). 
     
     
         18 . The pharmaceutical composition according to  claim 15 , wherein said pro-apoptotic protein member of the Bcl-2 family is any one of Bax, Bak, Bnip3, Nix/Bnip3L, Bid, Noxa, Puma and Bad. 
     
     
         19 . A method for treating, inhibiting, preventing, ameliorating or delaying the onset of a Bcl-2 over-expressing pathological disorder, said method comprises the step of administering to a subject in need thereof a therapeutically effective amount of at least one antagonist of a Bcl-2 prosurvival protein comprising ARTS or any fragment, peptide, analogues and derivatives thereof according to  claim 1 , or any composition comprising the same, wherein said fragment or peptide of ARTS comprises a BH3-like domain. 
     
     
         20 . (canceled) 
     
     
         21 . The method according to  claim 19 , further comprising the steps of:
 (a) determining the level of expression of at least one Bcl-2 prosurvival protein in at least one biological sample of said subject to obtain an expression value and;   (b) determining if the expression value obtained in step (a) is any one of, positive or negative with respect to a predetermined standard expression value or to an expression value of said Bcl-2 in a control sample;   
       Wherein a positive expression value of said Bcl-2 indicates that said subject is to be administered with said antagonist. 
     
     
         22 . The method according to  claim 21  comprising the step of:
 (a) determining the level of expression of at least one Bcl-2 prosurvival protein in at least one biological sample of said subject to obtain an expression value; 
 (b) determining if the expression value obtained in step (a) is any one of, positive or negative with respect to a predetermined standard expression value or to an expression value of said Bcl-2 in a control sample; and 
 (c) administering to a subject displaying a positive expression value of Bcl-2 as determined in step (b), a therapeutically effective amount of at least one antagonist of a Bcl-2 prosurvival protein comprising ARTS or any fragment, peptide, analogues and derivatives thereof or any composition comprising the same, wherein said fragment or peptide of ARTS comprises a BH3-like domain. 
 
     
     
         23 - 25 . (canceled) 
     
     
         26 . A combined composition comprising a therapeutically effective amount of:
 a. at least one antagonist of a Bcl-2 prosurvival protein comprising ARTS or any fragment, peptide, analogues and derivatives thereof, according to  claim 1 , wherein said fragment or peptide of ARTS comprises a BH3-like domain; and   b. any one of at least one BH3 mimetics compound and at least one pro-apoptotic member of the Bcl-2 family, wherein said BH3-mimetics compound is at least one of 4-[[2-(4-Chlorophenyl)-5,5-dimethyl-1-cyclohexen-1-yl]methyl]-1-piperazinyl]-N-[[4-[[(1R)-3-(4-morpholinyl)-1-[(phenylthio)methyl]propyl]amino]-3-[(trifluoromethyl)sulfonyl]phenyl]sulfonyl]benzamide (ABT-263), (R)-4-(4-((4′-chloro-[1,1′-biphenyl]-2-yl)methyl)piperazin-1-yl)-N-((4-((4-(dimethylamino)-1-(phenylthio)butan-2-yl)amino)-3-nitrophenyl)sulfonyl)benzamide (ABT-737), 1,1′,6,6′,7,7′-hexahydroxy-5,5′-diisopropyl-3,3′-dimethyl-[2,2′-binaphthalene]-8,8′-dicarbaldehyde (AT-101), (Z)-2-(2-((3,5-dimethyl-1H-pyrrol-2-yl)methylene)-3-methoxy-2H-pyrrol-5-yl)-1H-indole methanesulfonate (GX15-070), or DNA, d(P-thio)(T-C-T-C-C-C-A-G-C-G-T-G-C-G-C-C-A-T) (oblimersen sodium), and wherein said pro-apoptotic protein member of the Bcl-2 family is any one of Bax, Bak, Bnip3, Nix/Bnip3L, Bid, Noxa, Puma and Bad.   
     
     
         27 - 29 . (canceled) 
     
     
         30 . The combined composition according to  claim 26 , wherein said composition is a pharmaceutical composition for treating, inhibiting, preventing, ameliorating or delaying the onset of a Bcl-2 over-expressing pathological disorder, said composition optionally further comprises at least one pharmaceutically acceptable carrier, diluent or excipient. 
     
     
         31 . The method according to  claim 19 , for treating, inhibiting, preventing, ameliorating or delaying the onset of a Bcl-2 over-expressing pathological disorder, said method comprises the step of administering to a subject being treated with at least one BH3-mimetics compound, a therapeutically effective amount of at least one antagonist of a Bcl-2 prosurvival protein comprising ARTS or any fragment, peptide, analogues and derivatives thereof, wherein said fragment or peptide of ARTS comprises a BH3-like domain. 
     
     
         32 - 33 . (canceled) 
     
     
         34 . A kit comprising:
 (a) at least one antagonist of a Bcl-2 prosurvival protein comprising ARTS or any fragment, peptide, analogues and derivatives thereof, according to  claim 1 , optionally, in a first dosage form, wherein said fragment or peptide of ARTS comprises a BH3-like domain; and   (b) any one of at least one BH3 mimetics compound, at least one pro-apoptotic protein member of the Bcl-2 family and any combinations thereof, optionally, in a second dosage form.   
     
     
         35 . A prognostic method for determining the efficacy and assessing responsiveness of a mammalian subject to a BH3 mimetics treatment, said method comprises the steps of:
 (a) determining the level of expression of ARTS in at least one biological sample of said subject to obtain an expression value;   (b) determining if the expression value obtained in step (a) is any one of, positive or negative with respect to a predetermined standard expression value or to an expression value of ARTS in at least one control sample;   
       wherein a positive expression value of ARTS indicates that said subject belongs to a pre-established population associated with responsiveness to BH3 mimetics treatment. 
     
     
         36 . A method for determining a BH3 mimetics treatment regimen for a subject suffering from a Bcl-2 over expressing pathological disorder, the method comprises the steps of:
 (a) determining the level of expression of ARTS in at least one biological sample of said subject, to obtain an expression value; and   (b) determining if the expression value obtained in step (a) is any one of, positive or negative with respect to a predetermined standard expression value or to an expression value of ARTS in a control sample;   
       wherein a negative expression value of said ARTS indicates that at least one antagonist of a Bcl-2 prosurvival protein comprising ARTS or any fragment, peptide, analogues and derivatives thereof or any composition comprising the same is required in addition to said BH3 mimetics treatment for said subject, wherein said fragment or peptide of ARTS comprises a BH3-like domain. 
     
     
         37 . The method of  claim 36 , wherein said antagonist is administered before, after, simultaneously or any combinations thereof with any one of at least one BH3 mimetics compound, at least one pro-apoptotic protein member of the Bcl-2 family and any combinations thereof.